• Title/Summary/Keyword: Single strand DNA

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The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Lim Sang-Cheol;Rhim Tae-Jin
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.470-478
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    • 2005
  • The objective of present study was to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tort-butyl hydroperoxide(t-BHP), potent oxidizing agent to liver, for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Incubation with t-BHP increased glutamic oxaloacetic transaminase(GOT) and lactate dehydrogenase(LDH) acitivities and thiobarbituric acid reactive substances(TBARS) concentration but decreased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) reduction. Onion extracts at the concentration of 0.05 mg/ml decreased t-BHP-induced GOT and LDH activities. Onion extract at the concentration of 0.1 mg/ml increased t-BHP-induced MTT reduction. Onion extract at the concentration of 0.01 mg/ml decreased t-BHP-induced TBARS concentration. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, glutathione peroxidase(GSH-Px) and glutathione reductase(GSH-Rd) activities of hepatocytes were significantly decreased by t-BHP. Onion extracts at the concentration of 0.1 mg/ml prevented t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton reagents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition of lipid peroxidation.

Molecular Characteristics of Phytophthora katsurae Using PCR-SSCP Analysis (PCR-SSCP 분석에 의한 Phytophthora katsurae의 분자생물학적 특성)

  • Lee, Sun-Keun;Jang, Ha-Na;Lee, Dong-Hyeon;Lee, Sang-Hyun;Lee, Sang-Yong;Lee, Jong-Kyu
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.169-176
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    • 2011
  • Phytophthora katsurae is the fungus responsible for chestnut ink disease. The objectives of this study were to determine if a single-strand conformation polymorphism (SSCP) analysis of rDNA-ITS region, elongation factor 1 alpha gene and ${\beta}$-tubulin gene could be used for rapid identification and genetic diversity of P. katsurae, and to assess the potential use of the SSCP technique as a diagnostic tool for P. katsurae. Each regions amplified by PCR using primers designed to overlap the genus Phytophthora were characterized for the Phytophthora species. PCR products were denatured and electrophoresed for SSCP analysis. P. katsurae isolates showed an unique pattern in SSCP analysis and were easily distinguished from other Phytophthora species used as the control. This indicates that SSCP analysis is an useful technique for distinguishing Phytophthora species from genetically close relatives, and show that the SSCP analysis of each region is an efficient detection tool for P. katsurae. But PCR-SSCP analysis of single-gene may have difficulty in distinguishing P. katsurae from other Phytophthora species. Therefore, PCR-SSCP analysis of multi-genes can be useful for rapid and effective identification of P. katsurae.

Mutations of p53 Tumor Suppressor Gene in Human Lung Cancer Cell Lines (사람 폐암세포주에서 p53 종양억제유전자의 변이)

  • Hong, Weon-Seon;Hong, Seok-Il;Lee, Dong-Soon;Son, Young-Sook;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.6
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    • pp.653-658
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    • 1993
  • Background: Recent advancement of molecular genetics has revealed that malignant transformation of a cell may be a complex multistep process and this process is grouped, in general, into two distinct categories, activation of protooncogenes and inactivation of tumor suppressor genes. This study was focused on the mutation of p53 tumor suppressor gene, because p53 gene mutation is now generally accepted to be one of the most frequent genetic changes in a variety of human cancers. Although lung cancer is one of the common cancers in Korea, the genetic change in the carcinogenesis process is not yet known clearly. To investigate the role of p53 gene mutation in lung cancer, we examined the mutations of exon 4-8 of the p53 gene in humna lung cancer cell lines, because most of the mutations of p53 gene have been reported to develop in exon 4-8. Method: Genomic DNA was obtained by the digestion of proteinase K and the extraction by phenol-chloroform-ethanol method from two human pulmonary adenocarcinoma cell lines, PC-9 and PC-14, and one human small cell lung cancer cell line, H69. To detect the mutations of exon 4-8 of the p53 gene, polymerase chain reaction single-strand conformation polymorphism(PCR-SSCP) analysis was performed with the DNA extracted from the cells. Results: The mutation of p53 gene was present in all three cell lines tested. In PC-9, PC-14 and H69, the altered mobility was detected in exon 7, 7 and 5, respectively. Conclusion: These results suggest that p53 gene mutation plays an important role in certain steps of the carcinogenesis of human non-small cell and small cell lung cancer.

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Occurrence of Two Tobamovirus Diseases in Cucurbits and Control Measures in Korea

  • Park, Gug-Seoun
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.243-248
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    • 2001
  • Two Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV) and Zucchini green mottle mosaic virus (ZGMMV), occurred in Korea in 463 ha in 1998, 33.9 ha in 1999, and 44.2 ha in 2000. CGMMV was detected in watermelon, cucumber, oriental melon, and melon, whereas ZGMMV was mainly detected in zucchini squash. Thirty-six CGMMV isolates wee classified into three types by analysis of single strand cDNA conformational polymorphism (SSCP) of the coat protein gene. In a comparison of serological relationships among CGMMV, ZGMMV, and Kyuri green mottle mosaic virus (KGMMV), the three tobamoviruses specifically reacted with each homologous antibody in the double-antibody sandwich enzyme-linked immunosorbent assay and rapid imunofilter paper assay (RIPA), although ZGMMV and KGMMV were slightly biologcially similar. In a survey of the three tobamoviruses in cucurbitgrowing field in Korea by RIPA, CGMMV and ZGMMV were detected but KGMMV was not found in commercially growing cucurbit crops so far. Seed contamination ratio of CGMMV in bottle gourd seeds tested was 84%, while seed trasmission ratio from the virus-contaminated seeds was 2.0%. Soil transmission ratio was 0-3.5% in fields naturally infested with CGMMV or ZGMMV. Control measures of the virus diseases are roguing and sanitation. These suggest that it is important to rogue the first infected crops, which include the seed and soil, especially early in the season. This may be practicable to control the diseases because CGMMV and ZGMMV have a narrow host range restricted to cucurbitaceous crops.

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Effects of Rad51 on Survival of A549 Cells

  • Yu, Sha-Sha;Tu, Yi;Xu, Lin-Lin;Tao, Xue-Qin;Xu, Shan;Wang, Shan-Shan;Xiong, Yi-Feng;Mei, Jin-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.175-179
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    • 2015
  • Rad51, a key factor in the homologous recombination pathway for the DNA double-strand break repair, plays a vital role in genesis of non-small-cell lung cancer (NSCLC). In recent years, more and more studies indicate that high expression of Rad51 is of great relevance to resistance of NSCLC to chemotherapeutic agents and ionizing radiation. However, the underlying molecular mechanisms are poorly understood. In this study, we investigated the role of single Rad51 on cell viability in vitro. Our results show that depletion of endogenous Rad51 is sufficient to inhibit the growth of the A549 lung cancer cell line, by accumulating cells in G1 phase and inducing cell death. We conclude that independent Rad51 expression is critical to the survival of A549 cells and can be an independent prognostic factor in NSCLC patients.

Effect of Dioxin on the Change of Mitochondrial Inner Membrane Potential and the Induction of ROS (다이옥신이 미토콘드리아 내막의 전위차 변화 및 ROS 생성에 미치는 영향)

  • Cho, Il-Young;Sheen, Yhun-Yhong
    • Environmental Analysis Health and Toxicology
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    • v.24 no.1
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    • pp.33-41
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    • 2009
  • Among the toxicants in the environment dioxin-like compounds, including TCDD(2,3,7,8-Tetrachlorodibenzo-p-Dioxin), are well known as carcinogen and teratogen. TCDD the most toxic of these compounds, may result in a wide variety of adverse health effects in humans and environment, including carconogenesis, hepatotoxicity, teratogenesis, and immunotoxicity. Also TCDD increases superoxide, peroxide radicals and induces oxidative stress that leads to breakage of DNA single-strand and mitochondrial dysfunction. Recently, there have been reports that persistent organic pollutants(POPs) may be causing metabolic disease through mitochondrial toxicity. In order to examine if dioxin brings about toxicity on mitochondria directly, we measured the change of the mitochondrial membrane potential after exposure to TCDD using JC-1 dye. After short time exposure of dioxin, mitochondrial depolarization was observed but it recovered to the control level immediately. This TCDD effect on mitochondrial membrane potential was not correlated either to the production of reactive oxygen species(ROS) or extracellular $Ca^{2+}$ by TCDD. Less than 2 hours exposure of TCDD did not show any change in ROS production but 0.25 nM TCDD for 48 hours or 0.5 nM TCDD for 12 hours exposure did increase in ROS production. Under these conditions of ROS production by TCDD, no changes in the mitochondrial membrane potential by TCDD was observed.

Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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An Overview of Different Techniques on the Microbial Community Structure, and Functional Diversity of Plant Growth Promoting Bacteria

  • Kim, Kiyoon;Islam, Rashedul;Benson, Abitha;Joe, Manoharan Melvin;Denver, Walitang;Chanratan, Mak;Chatterjee, Poulami;Kang, Yeongyeong;Sa, Tongmin
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.2
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    • pp.144-156
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    • 2016
  • Soil is a dynamic biological system, in which it is difficult to determine the composition of microbial communities. Knowledge of microbial diversity and function in soils are limited because of the taxonomic and methodological limitations associated with studying the organisms. In this review, approaches to measure microbial diversity in soil were discussed. Research on soil microbes can be categorized as structural diversity, functional diversity and genetic diversity studies, and these include cultivation based and cultivation independent methods. Cultivation independent technique to evaluate soil structural diversity include different techniques such as Phospholipid Fatty Acids (PLFA) and Fatty Acid Methyl Ester (FAME) analysis. Carbon source utilization pattern of soil microorganisms by Community Level Physiological Profiling (CLPP), catabolic responses by Substrate Induced Respiration technique (SIR) and soil microbial enzyme activities are discussed. Genetic diversity of soil microorganisms using molecular techniques such as 16S rDNA analysis Denaturing Gradient Gel Electrophoresis (DGGE) / Temperature Gradient Gel Electrophoresis (TGGE), Terminal Restriction Fragment Length Polymorphism (T-RFLP), Single Strand Conformation Polymorphism (SSCP), Restriction Fragment Length Polymorphism (RFLP) / Amplified Ribosomal DNA Restriction Analysis (ARDRA) and Ribosomal Intergenic Spacer Analysis (RISA) are also discussed. The chapter ends with a final conclusion on the advantages and disadvantages of different techniques and advances in molecular techniques to study the soil microbial diversity.

Association between Mismatch Repair Gene MSH3 codons 1036 and 222 Polymorphisms and Sporadic Prostate Cancer in the Iranian Population

  • Jafary, Fariba;Salehi, Mansoor;Sedghi, Maryam;Nouri, Nayereh;Jafary, Farzaneh;Sadeghi, Farzaneh;Motamedi, Shima;Talebi, Maede
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6055-6057
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    • 2012
  • The mismatch repair system (MMR) is a post-replicative DNA repair mechanism whose defects can lead to cancer. The MSH3 protein is an essential component of the system. We postulated that MSH3 gene polymorphisms might therefore be associated with prostate cancer (PC). We studied MSH3 codon 222 and MSH3 codon 1036 polymorphisms in a group of Iranian sporadic PC patients. A total of 60 controls and 18 patients were assessed using the polymerase chain reaction and single strand conformational polymorphism. For comparing the genotype frequencies of patients and controls the chi-square test was applied. The obtained result indicated that there was significantly association between G/A genotype of MSH3 codon 222 and G/G genotype of MSH3 codon 1036 with an increased PC risk (P=0.012 and P=0.02 respectively). Our results demonstrated that MSH3 codon 222 and MSH3 codon 1036 polymorphisms may be risk factors for sporadic prostate cancer in the Iranian population.

Responses of Various Biomarkers in Common Carp (Cyprinus carpio) Exposed to Benzo[k]fluoranthene

  • Kim, Woo-Keun;Kim, Ja-Hyun;Yeom, Dong-Hyuk;Lee, Sung-Kyu
    • Korean Journal of Ecology and Environment
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    • v.41 no.3
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    • pp.331-337
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    • 2008
  • Polycyclic aromatic hydrocarbons (PAHs) derived from leakage of fossil fuels and incomplete combustion of organic materials have been considered as harmful contaminants in environments. This study evaluated the effect of benzo[k]fluoranthene (BkF), one of the PAHs, using the multiple biomarkers and applied the integration model with those biomarker responses. After 10 days of the exposure at the measured concentrations of BkF (6, 25, and 45 ${\mu}g\;L^{-1}$), the changes of the four biomarkers, that is, 7-ethoxyresorufin-O-deethylase (EROD), DNA single-strand breaks (Comet), acetylcholinesterase (AChE) and vitellogenin (VTG) in the common carp (Cyprinus carpio) were observed. The standardized values of four biomarker responses were computed and integrated as star plots, representing Integrated Biomarker Respnse (IBR) values. DNA damage was induced in a dose-dependent manner, and increased significantly compared with that in the control. EROD and VTG levels were significantly elevated at low concentrations of BkF. On the other hand, AChE activities were not altered by BkF. IBR values increased as the exposure concentrations increased. Thus, the metabolic, endocrine and genetic changes of the biomarker responses in the common carp exposed to BkF should be considered in the case of the ecological risk assessment of the BkF in fish and it can be used as a biomonitoring tool in aquatic ecosystems. In addition, star plots can be used as a useful analysis tool in multibiomarker integration approach.